View source for Marek: Freeze-down/Thaw

==Overview==

This short protocol describes how to freeze down and thaw (or bring up) mammalian cells (e.g. HeLa, KEK293, CHO, Jurkat T cells).

==Materials==
List reagents, supplies and equipment necessary to perform the protocol here.

*10-50 ml centrifuge tube 
*DMSO, Tissue culture (TC)-tested (e.g. Sigma, D2650)
*Cell culture medium (depends on cell line/cells)
*Trypsin-EDTA (e.g. from [http://www.paa.com PAA], L11-004) - for adeherent cells
*Freeze-down medium
**6% DMSO
**Cell culture medium (you can use 100% fetal calf serum (FCS) instead of cell culture medium)
*Basic TC centriguge (0 - 2500 rpm)
*Freeze-down box (this could be simply made from a small styropore box and cotton or from two styropore racks from 15-ml centrifuge tubes )

==Procedure==
#Step 1
#Step 2
#*Step 2 has some additional information that goes with it.  i.e. Keep at 4&deg;C.
#Step 3
##Step 3 has multiple sub-steps within it.
##Enumerate each of those.

==Notes==
#List troubleshooting tips here.  
#You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
#Anecdotal observations that might be of use to others can also be posted here.  

Please sign your name to your note by adding <font face="courier"><nowiki>'''*~~~~''':</nowiki></font> to the beginning of your tip.



==Contact==
*Who has experience with this protocol?

or instead, [[Talk:{{PAGENAME}}|discuss this protocol]]. 

<!-- You can tag this protocol with various categories.  See the [[Categories]] page for more information. -->
[[Category:Mammalian cell culture]]
[[Category:In vitro]]
[[Category:Protocol]]